[Hypoxia reduces the proliferation and occludin expression of primary sertoli cells].

OBJECTIVE: To investigate the effect of hypoxia on the proliferation and occludin expression of primary rat Sertoli METHODS: We constructed a primary Sertoli cell system by two-step enzymatic digestion in 18 -22 days old Wistar rats and identified it by oil red O and immunofluorescence methods. We randomly divided the Sertoli cells into five groups to be cultured in oxygen at the concentrations of 20%, 15%, 10%, 5% and 1%, respectively, for 6, 12, 24, 48 and 72 hours. We detected the proliferation of the Sertoli cells by CCK-8 assay, determined the expression of occludin by Western blot, and analyzed the differences among the five groups. RESULTS: Oil red O staining revealed red lipid droplets in the cytoplasm of the Sertoli cells, and immunofluorescence showed the positive expression of the FasL protein, with the purity of Sertoli cells over 95% in vitro. Compared with the 20% normoxic group, the proliferation of the Sertoli cells was gradually reduced in the 15% and 10% hypoxia groups, and significantly declined in the 5% and 1% groups (P < 0.01). At 12 hours, the expression of occludin began to decrease with the prolonging of time and reduction of oxygen concentration (P < 0.01). CONCLUSION: Hypoxia suppresses the proliferation of Sertoli cells and reduces the expression of occludin. It could be inferred that hypoxia could damage the integrity of blood-testis barrier and spermatogenesis of the testis.

[High temperature reduces the proliferation of and occludin expression in rat Sertoli cells in vitro].

OBJECTIVE: To explore the action mechanisms of temperature in male infertility or subfertility by observing the effects of different temperatures on the proliferation of and occludin (OCLN) expression in rat Sertoli cells in vitro. METHODS: We isolated Sertoli cells from the testis of male Wistar rats, and performed oil red O staining and immunohistochemistry to identify their FasL. We cultured the Sertoli cells at 34 degrees C (control group) and at 35, 36, 37, 38 and 39 degrees C (experimental groups) for 4 days. Then we measured their proliferation by CCK-8 assay, observed their morphology and structure by hematoxylin-eosin staining, and determined their OCLN expression level by Western blotting and immunofluorescence. RESULTS: The purity of the isolated Sertoli cells was (96.20 +/- 1.95)%. CCK-8 assay indicated that the proliferation of the Sertoli cells was increased between 34 and 36 degrees C, and decreased at 36-39 degrees C. The pyknotic nuclei and fragmentation of the Sertoli cells were more obvious at > 36 degrees C. Western blot and immunofluorescence showed the highest level of OCLN expression at 36 degrees C, which, however, decreased while the temperature rose above 36 degrees C (P < 0. 01). CONCLUSION: High temperature (> 36 degrees C) inhibited the proliferation of rat Sertoli cells in vitro, and decreased the expression of OCLN, which suggests that a higher temperature above 36 degrees C may reduce male fertility by affecting the proliferation of Sertoli cells and integrity of the tight junction among Sertoli cells or Sertoli cells and other cells.

[Partial least-squares regression analysis for simultaneous spectrophotometric determination of Fe3+ and Al3+].

Adding hexodecyltrimethylammonium bromide(CTMAB) to improve solubility and delicacy, and using chromazurol S as a chromogenic reagent, and acetone as a stabilizing reagent, partial least-squares regression(PLS) was used to determine simultaneously trace amounts of Fe(III) and Al(III) in sythetic samples. Considering the stability of the rays, and the absorption properties of the two ingredients, seven wavelengths between 610 and 670 nm were selected for data collection. As a comparison, the methods of CPA and PLS were used respectively for analysing the data obtained from the experiment, and the results suggested that the latter was much better.

[Treatment of wastewater containing Cr(VI) by LDH synthesizing in situ].

The objective of this work was to investigate the efficiency and factors impacting of removal Cr(VI) from wastewater by layer double hydroxide synthesizing in situ. Principle of the method may be described as follow: Mg2+ and Al3+ hydrolysis and forms Mg/Al-LDH by adding Mg2+, Al3+ and NaOH into wastewater containing Cr(VI), Cr(VI) anions are selectively intercalated into interlayer of LDH to balance positive structural charge. While Mg2+ and Al3+ co-precipitates and forms LDH, the Cr(VI) in wastewater is removal by settle of LDH synthesizing in situ, which are confirmed by analysis of X-ray diffraction on settle and chemical analysis on aqueous. The results indicate that factors of impacting on efficiency of removal Cr(VI) include in amount of adding Mg2+ and Al3+, Mg/Al ratio, pH and concentration of Cr(VI) in wastewater. The maximal removal efficiency of Cr(VI) can be reached when pH values are between 8.5 and 9, and Mg/Al ratios are between 1:1 and 2:1, meanwhile, Mg and Al added can be taken good use of. This technology has present extraordinary efficiency of wastewater treatment.